Non Technical Summary Anthocyanin pigments and polyphenolics provide several health benefits because of their antioxidant properties. There is need for compositional information on their concentrations, distribution, and antioxidant properties. The purpose of this project is to identify and measure the anthocyanin pigments and polyphenolics in fruit and vegetable processing wastes and potential new crops.

Project Methods Sample Materials Fruit and vegetable samples will be obtained from the OSU Horticulture Department, the USDA Northwest Center for Small Fruit Research, the Corvallis USDA Germplasm Repository, and the USDA/ARS Research and Extension Center, Prosser, WA. Samples of fruit, seeds, and juice processing waste streams will be obtained from industrial collaborators. Extraction and Isolation of Anthocyanins and Polyphenolics Cryogenic milling, acetone extraction, chloroform partition, and solid-phase extraction with C-18 resin will be used. Identification of Anthocyanins and Polyphenolics HPLC Analyses- Analytical and semi-preparative reverse phase HPLC is the primary analytical tool for separation and isolation of individual compounds. These instruments are equipped with u.v.-visible diode array detectors which is very useful in identifying compounds. Mass Spectroscopy (MS)- The Food Science Department has a Perkin Elmer Sciex III triple-quadrupole ionspray mass spectrometer
that we will be using for HPLC-MS for anthocyanins and polyphenolics. NMR will be done on a fee basis at the Department of Biochemistry and Biophysics, OSU. Measurement of Anti-Oxidant Activity Oxygen-Radical Absorbance Capacity (ORAC) and Ferric Reducing Antioxidant Potential (FRAP) will be conducted at the Linus Pauling Institute. Measurements will be made on extracts, isolated fractions, and individual compounds. Colorant Properties Quantitative Pigment Measurements- Total anthocyanins will be measured by the pH differential method; additional spectral measurements will include indices for polymeric color, color density and browning. Color Measurement- A Hunter ColorQuest colorimeter will be used for measuring CIE L*a*b* values, hue angle and chroma of pigment extracts. Processing Trials The Food Science and Technology Department pilot plant is well-equipped for conducting juice processing trials and preparing aqueous natural colorant preparations, e.g., Hydraulic and Willmes Bag
presses, Stephan vertical cutter mixer, APV-Crepaco heat-exchanger, Westfalia continuous centrifuge, Strassburger plate filtration unit, Alfa-Laval Centritherm evaporator. Data Analysis All experiments will be replicated and analyses will be done in duplicate. The data will be subjected to ANOVA and other statistical procedures to determine level of significance and standard error.

Progress 05/01/03 to 09/20/07

OutputsThe project team organized the symposium "Color Quality of Fresh and Processed Foods" (Cathy Culver and Ron Wrolstad, co-organizers), which was held at the American Chemical Society meeting in Atlanta, GA, March 26-30, 2006. The monograph is being completed, and should be published as an ACS Symposium Series Book sometime in 2007 (it generally takes a year or so after the symposium to publish).

ImpactsInterest in anthocyanin pigments has continued to intensify because of their possible health benefits in reducing the risk of chronic diseases such as coronary heart disease, cancer and stroke. The conference provided a forum for speakers to present information for a wide range of food commodities.

Publications

No publications reported this period

Progress 01/01/05 to 12/31/05

OutputsINFLUENCE OF VARIETY, GROWING REGION, WINERY AND AGING ON THE ANTHOCYANIN PIGMENT COMPOSITION OF RED WINES. We are analyzing the anthocyanin pigment composition of 44 varietal pure commercial wines from California, Oregon and Washington. The major objective is to determine whether varietal origin of red wines can be determined from analysis of their anthocyanin pigments. In addition, we want to determine the influence of growing regions, winery, and aging on anthocyanin pigment composition. HPLC-MS analyses have been completed at zero and six month's storage time. Data analysis is in progress. STRAWBERRY COLOR QUALITY. CIEL*a*b* indices, spectrophotometric measurements and HPLC of anthocyanin pigments are being used to measure the color quality of fresh and processed strawberries. The color of fresh strawberries is dramatically affected by processing. While some pigment degradation occurs with freezing, physical changes, which are manifested by drip loss, are largely
responsible for deterioration of appearance. Canning and manufacture into jam, juice, and wine results in marked color degradation. Pigment concentration, browning reactions, and co-pigmentation can have an impact on the color quality of these products. Reaction of anthocyanins with enzymes, ascorbic acid, acetaldehyde and other reactive compounds account for much of the pigment degradation. L*, hue angle and chroma are very useful indices for monitoring color change, and they are complementary to measurement of total anthocyanins, polymeric color and anthocyanin pigment profiles.

ImpactsAOAC Collaborative Study, Determination of Total Monomeric Anthocyanin Pigment Content of Fruit Juices, Beverages, Natural colorants and Wines by the pH Differential Method. Interest in anthocyanin pigments has intensified because of their possible health benefits in reducing the risk of chronic diseases such as coronary heart disease, cancer and stroke. There has been no standardized method for measuring anthocyanin pigment content. Representatives of the fruit juice, functional foods and nutraceutical industries asked our laboratory to organize and conduct a collaborative study for measuring total anthocyanins. Planning, pre-testing, conducting the study, and AOAC review took several years. AOAC approved the study as Official First Action, and the manuscript was published in 2005. Color Quality: We are working collaboratively with Yanyun Zhao and Chad Finn in evaluating the color quality of fresh and processed strawberries. Color and appearance are extremely
important, as clients and consumers will only accept a product if the color is "right". We have organized a three-day Symposium, Color Quality of Fresh and Processed Foods, for the spring 2006 ACS meeting that will be published as an ACS monograph. There are 36 speakers covering a wide range of food commodities.

OutputsPOTENTIAL NEW CROPS, COMPOSITION AND QUALITY: Anthocyanins and phenolics of blue honeysuckle (Lonicera caerulea L.) were characterized and quantified by HPLC-DAD and ESMS. Cyanidin-3-glucoside was the major peak, with the additional minor peaks being cyanidin-3,5-diglucoside, cyanidin-3-rutinoside, pelargonidin-3-glucoside, peonidin-3-glucoside, and peonidin-3-rutinoside. Four polyphenolics were identified as chlorogenic acid, neochlorogenic acid, quercetin-3-rutinoside, and quercetin-3-glucoside. Total anthocyanins (n = 11) ranged from 116-593 mg/100g, total phenolics from 427-1140 mg GAE/100 g, hydroxycinnamates from 30.4 to 156.2 mg/100g, flavonols from 12.6 to 32.8 mg/100g, and ORAC from 21,840-10,370 micromol Trolox equivalents/g. The total anthocyanins and total phenolics of PNW wild and cultivated Huckleberry species were evaluated. Total anthocyanin content ranged from 101 to 400 mg/100g, and total phenolics from 367 to 1286 mg/100g. Significant variations
were observed among the different V. membranaceum, V. ovalifolium, and V. deliciosum populations. The anthocyanin pigment profiles of wild V. membranaceum, wild V. ovalifolium, and V. corymbosum Rubel were determined. The chromatograms of V. membranaceum, V. ovalifolium, and Rubel were distinctly different in the amounts of delphinidin, cyanidin, and malvidin glycosides. Lingonberry (Vaccinium vitis-idaea) samples (n = 11) were analyzed with the following results: Total anthocyanins ranged from 12-44 mg/100g with a mean of 33; total phenolics ranged from 532-984 GAE mg/100g with a mean of 706; pH ranged from 3.52-3.67; TA ranged from 0.55-1.46 g citric acid/100 mL, mean of 0.87. ORAC ranged from 18-41 micromol Trolox eq/g, mean of 28. Anthocyanin pigment profiles were very similar with cyanidin-3-galactoside being the major pigment (70-85.5% of total peak area), and cyanidin-3-glucoside (4.4-16.7%) and cyanidin-3-arabinoside (1.7-12.9%) as secondary pigments. Chokeberry (Aronia
melanocarpa) samples (n = 6) were analyzed with the following results: Total anthocyanin pigments ranged from 380-497 mg/100 g, mean of 430; total phenolics ranged from 159-183 mg GAE/100g. mean of 171; pH ranged from 3.51-3.67; TA ranged from 0.35-0.57 mg citric acid/100 mL, mean of 0.45; ORAC ranged from 30-183 micromol Trolox eq/g, mean of 88. Anthocyanin pigments showed similar profiles: cyanidin-3-galactoside (67.1-68.4 % of total peak area), cyanidin -3-arabinoside (23.6-24.9%), cyanidin-3-glucoside 2.6-3.0%), cyanidin -3-xyloside (2.9-3.4%). INFLUENCE OF VARIETY, GROWING REGION, WINERY AND AGING ON THE ANTHOCYANIN PIGMENT COMPOSITION OF RED WINES. We are analyzing the anthocyanin pigment composition of 44 varietal pure commercial wines from California, Oregon and Washington. The major objective of this study to determine whether varietal origin of red wines can be determined from analysis of their anthocyanin pigments. In addition, we want to determine the influence of growing
regions, winery, and aging on anthocyanin pigment composition. The anthocyanin profiles have been determined by HPLC, with peak identities confirmed by LC-MS. Data analysis is in progress.

ImpactsFood processors and nutritionist are interested in developing fruit and vegetable varieties that contain higher amounts of dietary antioxidants, in addition to other desirable quality attributes and horticultural properties. Working with plant breeders, we have analyzed the anthocyanin pigment, polyphenolic composition, and antioxidant properties of new selections of several fruits and vegetables. Some of the experimental blackberry selections were higher in total anthocyanins and total phenolics than the common commercial varieties showing the potential for obtaining new cultivars with high pigment/phenolic content through classical plant breeding. The development of potatoes with red and purple flesh pigmentation offers the possibility of new product line extensions, in addition to having potatoes with higher levels of dietary antioxidants. One Oregon-based company has purchased exclusive rights to a red-fleshed potato that is being used for potato chips that are
burgundy-colored and also have high levels of dietary antioxidants. We are working collaboratively with horticulturists to evaluate plant materials that have potential for being new commercial crops in the Pacific Northwest. We have analyzed Pacific Northwest huckleberries that have been collected from the wild and grown in cultivated plots. The fruits are very rich in anthocyanin pigments and dietary antioxidants. Blue Honeysuckle was collected in Japan, Russia and China. Selections grown in Corvallis ripen early (May-June) and are very rich in anthocyanin pigments and dietary antioxidants.

OutputsBLACKBERRIES: Total anthocyanins, total phenolics and antioxidant properties for 11 blackberry cultivars were determined. Total anthocyanins for the 11 cultivars ranged from 131-256 mg/100 g, total phenolics from 682-1056 mg GAE/100g and ORAC from 37.6-75.5 micro-mol TE/g. The influence of ripening on the polyphenolic composition of Evergreen and Marion blackberries was investigated with total anthocyanins increasing from 74.7 to 317 mg/100g from partial ripe to overripe for Marions, and from 69.9 to 164 for Evergreen. Total phenolics did not show marked change with maturity. Antioxidant activities did not show the marked incrase with ripening that anthocyanins exhibited. BLUEBERRIES: The anthocyanins and polyphenolics of wild and cultivated Pacific Northwest huckleberries were investigated. Total anthocyanins ranged from 101 to 400 mg/100g, and total phenolics from 367 to 1286 mg/100g. A comparison of cultivated V. membranaceum and V. ovatum showed that the smaller
berry size of V. ovatum resulted in much higher anthocyanins, total phenolics and antioxidant properties. CHERRIES: Anthocyanins, polyphenolics and antioxidant properties of 1 sour cherry and 3 sweet cherry cultivars were investigated. Bing cherries were highest in anthocyanins while Montmorency was highest in total phenolics and antioxidant properties. Total phenolics and anthocyanins were concentrated in the skin. Changes during frozen storage, canning and brining were monitored. Hydroxycinnamates were greatly affected by processing and storage while flavonol glycosides were quite stable. Half of the anthocyanins and polyphenolics were transferred to the syrup with canning, and nearly all were transferred to brine during brining. PEARS: The polyphenolic composition and antioxidant properties of the peel and flesh of 6 varieties of pears were analyzed. The flesh was very low in polyphenolics while the peels had high concentrations. Chlorogenic acid was the major polyphenolic in both
peel and flesh for all cultivars. The effect of ripening on Starkrimson and Bartlett polyphenolics showed little quantitative difference between the two stages. Antioxidant properties had a high correlation with total phenolics. POTATOES: Total anthocyanins ranged from 6.9-35 mg/100g in red fleshed, and 5.5-17.1 mg/100g in purple fleshed breeding clones. Acylated glycosides of pelargonidin predominated in red-fleshed clones while acylated petunidin and pelargonidin-glycosides predominated in purple-fleshed clones. Antioxidant properties (ORAC and FRAP) were 2-3 times higher than white-fleshed potatoes. BLUE HONEYSUCKLE: Anthocyanins, total phenolics and antioxidant properties of 10 blue honeysuckle (Lonicera caerulea L.) genotypes were analyzed, and the 6 anthocyanins identified. AOAC COLLABORATIVE STUDY: We conducted a collaborative study "Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants and wines by the pH differential
method". Agreement between the 11 participating collaborators was excellent and we have recommended that the method be adopted Official First Action by AOAC.

ImpactsFood processors and nutritionist are interested in developing fruit and vegetable varieties that contain higher amounts of dietary antioxidants, in addition to other desirable quality attributes and horticultural properties. Working with plant breeders, we have analyzed the anthocyanin pigment, polyphenolic composition, and antioxidant properties of new selections of several fruits and vegetables. Some of the experimental blackberry selections were higher in total anthocyanins and total phenolics than the common commercial varieties showing the potential for obtaining new cultivars with high pigment/phenolic content through classical plant breeding. The development of potatoes with red and purple flesh pigmentation offers the possibility of new product line extensions, in addition to having potatoes with higher levels of dietary antioxidants. One Oregon-based company has purchased exclusive rights to a red-fleshed potato that is being used for potato chips that are
burgundy-colored and also have high levels of dietary antioxidants. We are working collaboratively with horticulturists to evaluate plant materials that have potential for being new commercial crops in the Pacific Northwest. We have analyzed Pacific Northwest huckleberries that have been collected from the wild and grown in cultivated plots. The fruits are very rich in anthocyanin pigments and dietary antioxidants. Blue Honeysuckle was collected in Japan, Russia and China. Selections grown in Corvallis ripen early (May-June) and are very rich in anthocyanin pigments and dietary antioxidants.

Publications

LEE, J., C.E. FINN, and R.E. WROLSTAD. 2003. Anthocyanin pigment and total phenolic content of three Vaccinium species native to the Pacific Northwest of North America. Accepted for publication in HortScience.

CHAOVANALIKIT, A. and R.E. WROLSTAD. 2003. Total anthocyanins and total phenolics of fresh and processed cherries and their antioxidant properties. Accepted for publication in J. Food Sci.

Progress 04/01/97 to 03/31/02

OutputsThis project has emphasized the anthocyanin pigment and polyphenolic composition of fruits and vegetables as impacted by cultivar, processing and post-harvest storage. Antioxidant properties are of particular interest because of their possible relationship to health benefits. Utilization of juice processing wastes as sources for nutraceuticals and natural colorants is given attention, as is color quality of juices and concentrates. Natural colorants with superior properties of hue and stability were developed from radishes and red-fleshed potatoes. Potatoes- We have determined the anthocyanin pigment composition and antioxidant activities of several potato cultivars. Potato chips made from these selections have attractive colors, high anthocyanin content, and high antioxidant activities. Blueberries- Total anthocyanins, total phenolics, and antioxidant capacities for 30 Vaccinium, 37 Rubus, and 40 Ribes genotypes were determined. Processing wastes from blueberry juice
manufacture are a good source for natural colorants and nutraceuticals, being high in anthocyanin pigments, polyphenolics, and antioxidant activity. Since the bioactive compounds are concentrated in the skins, extracting the skins can give a rich colorant/nutraceutical preparation. Recovery of anthocyanins from blueberry skins can be increased by 40 percent with enzyme treatment. Blackberries- Blackberries seeds account for 5 percent of berry weight and are rich in ellagic acid derivatives and are also high in antioxidant activity. Cyanidin-3-dioxalyl-glucoside was identified as a novel zwitterionic anthocyanin pigment in blackberries. Apples-Total phenolics, total anthocyanins and antioxidant properties of Red Delicious, Granny Smith, and Fuji apples were determined. Antioxidant activity is primarily in the aqueous fraction and believed to be due to polyphenolics. Red Delicious is highest in total phenolics and antioxidant levels, Granny Smith next, and Fuji lowest. Apple extracts
and apple polyphenolics added to human plasma significantly protected endogenous antioxidants from oxidation and prevented lipid hydroperoxide formation. Pears- The antioxidant activities and polyphenolics in peel and flesh of the following cultivars were determined: Bartlett, Star crimson, Anjou, Red Anjou, Bosc and Comice. Cherries-Total anthocyanin pigments, total phenolics, and antioxidant activities for extracts of peel, flesh and pits of Bing, Royal Anne, Ranier and Montmorency cherries were determined. Total phenolics has a high correlation with antioxidant activities. Bing cherries showed substantial anthocyanin pigment degradation during frozen storage. Antioxidant levels were high in canned cherries, but 50 percent of the total phenolics and antioxidant activity is transferred from the fruit to the syrup. Anthocyanin Pigment Properties- The color and antioxidant properties of several purified cyanidin-based anthocyanin pigments were determined. The number of sugar residues,
position of glycosidic substitution and acylation with cinnamic acids had a marked effect on color properties, visual detection thresholds, hydration constants and in vitro antioxidant activities.

ImpactsThis project has provided plant breeders, food processors and nutritionists with information that should have a positive impact on human nutrition and health. Anthocyanin pigments and polyphenolics are natural antioxidants, which are believed to play a major role in the health benefits provided by fruits and vegetables. We identified and measured the concentrations of these compounds in apples, cherries, blueberries, blackberries and potatoes, and also measured their antioxidant activities. We also investigated the effects of variety, processing and post-harvest storage on these compounds. We have developed natural colorants from radishes and red-fleshed potatoes that exhibit superior color and stability properties.

OutputsResearch in year 2001 emphasized anthocyanin pigment and polyphenolic composition of fruits and vegetables as impacted by cultivar, processing and post-harvest storage. Commodities investigated included apples, cherries, blueberries, blackberries, carrots and potatoes. Antioxidant properties are of particular interest because of their possible relationship to health benefits. Utilization of juice processing wastes as sources for nutraceuticals and natural colorants is given attention, as is color quality of juices and concentrates. KEYFINDINGS: 1) Monitored changes in anthocyanins and polyphenolics during processing of blueberries into juice and concentrate. Press-cake residue contains 42-55 percent of fresh fruit anthocyanins and 16-20 percent of polyphenolics. 2) Determined the polyphenolic composition of Evergreen and Marion Blackberries. Marionberries are higher than Evergreen in total anthocyanins and total phenolics while Evergreen are higher in total flavonols,
ellagic acid forms and epicatechin. Seeds account for 5 percent of fruit weight and are rich in ellagic acid derivatives. 3) Identified a novel zwitterionic anthocyanin, cyanidin 3-dioxalyl-glucoside, as a minor pigment in blackberries. 4) Determined the total anthocyanin pigments, total phenolics, and antioxidant activities for extracts of peel, flesh and pits for Bing, Royal Anne, Ranier and Montmorency cherries. Anthocyanins, phenolics and antioxidant activities are highest in the skin. Bing and Montmorency are highest in anthocyanin. Montmorency is highest in total phenolics, with Royal Anne being slightly higher than Bing. Ranier is lowest. Total phenolics has a high correlation with antioxidant activities as measured by ORAC and FRAP. 5) Determined the color and antioxidant properties of several purified cyanidin-based anthocyanin pigments. The number of sugar residues, position of glycosidic substitution and acylation of sugar residues with cinnamic acids had a marked effect on
color properties, visual detection thresholds, hydration constants and in vitro antioxidant activities. Mixtures of anthocyanins had a synergistic effect on antioxidant activity. 6) Determined the total phenolics, total anthocyanins and antioxidant properties of Red Delicious, Granny Smith, and Fuji apples. Antioxidant activity is primarily in the aqueous fraction and believed to be due to polyphenolics. Red Delicious is highest in total phenolics and antioxidant levels, Granny Smith next, and Fuji lowest. ORAC values are higher in peel than flesh. Red Delicious peel has the highest ORAC value, presumably because of anthocyanin pigment content. There is no significant difference in ORAC between MCP-treated and control samples.7) Determined total anthocyanins, total phenolics, HPLC anthocyanin pigment profiles, and antioxidant activities of several potato cultivars. PATENTS: Wrolstad, R. E. & L. E. Rodriguez-Saona. Natural colorant from potato extract. Patent No. US 6,180,154 B1, Jan.
30, 2001.Wrolstad, R. E. & Ling Wen. Natural antibrowning and antioxidant compositions and method for making the same. Patent No.: US 6,224,926 B1; May 1, 2001.

ImpactsProject 422 provided information to plant breeders, food processors and nutritionists, which should have a positive impact on human nutrition and health. Anthocyanin pigments and polyphenolics are natural antioxidants, which are believed to play a major role in the health benefits provided by fruits and vegetables. We identified and measured the concentrations of these compounds in several fruits and vegetables, and also measured their antioxidant activities. We also investigated the effects of variety, processing and post-harvest storage on these properties.

OutputsGLYCOSIDASE ACTIVITY OF COMMERCIAL JUICE PROCESSING ENZYMES: Screening methods developed in our laboratory showed that 21 out of 28 commercial enzyme preparations for fruit juice processing contained (beta)-galactosidase activity. Pilot plant experiments were conducted using two enzyme preparations to determine if anthocyanin pigments were destroyed by the enzymes during cranberry juice processing. Significant losses of cyanidin-3-(beta)-galactoside occurred. APPLE JUICE PHENOLICS: The phenolic composition of the NFPA '93 authentic apple juice samples were analyzed by HPLC. Samples were qualitatively similar, but showed wide differences in quantitative amounts. Concentrations were in the order as measured in the '92 NFPA sample set. Arbutin was detected in 13 of the 26 samples. NATURAL COLORANTS FOR BRINED CHERRIES: Anthocyanin pigments from radish peels were extracted, purified, and characterized by HPLC / UV-visible spectra. The major pigment is
pelargonidin-3-sophoroside-5-glucoside acylated with p-coumaric and/or caffeic acids. Concentration is 150 mg/100 g. Brined cherries were processed into maraschino cherries, using radish extract as a colorant. Color was very similar to cherries colored with FDC Red No. 40.

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Publications

Progress 01/01/92 to 12/30/92

OutputsGLYCOSIDASE ACTIVITY OF COMMERCIAL JUICE PROCESSING ENZYMES: Methods have been developed for measuring the anthocyanin pigment degradative activity of enzymes. Twenty-eight commercial enzyme preparations were screened with 21 of the preparations exhibiting (beta)-galactosidase activity (from 10-98% destruction of cyanidin-3-galactoside). None of the preparations exhibited arabinosidase activity. The methods permit the selection of enzymes which will optimize color quality of cranberry juice. APPLE JUICE PHENOLICS: Concentrations of phenolics (chlorogenic acid, caffeic acid, p-coumaric acid, catechin, epicatechin, two procyanidins, phloridzin, rutin and hydroxymethylfurfural) have been determined in 34 authentic apple juice samples. This data will become part of the authentic apple juice database being developed by NFPA and FDA. CANTALOUPE JUICE PROCESSING: Cantaloupe juice has been concentrated into clarified and pulpy juice concentrates using ion-exchange and direct
osmotic concentration technologies. Composition and sensory quality are being evaluated. Cation exchange treatment reduces browning and prevents haze/sediment formation.

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Publications

Progress 01/01/91 to 12/30/91

OutputsRED RASPBERRY JUICE COMPOSITION: Pilot-plant processed juices (n = 46) were analyzed, sample differences including cultivar, processing method, maturity, geographic origin, mold contamination and harvesting method. The mean Brix (9.95 degree) is lower than the proposed USA standard (10.5 degree). Sorbitol is present in many samples at levels < 0.5% of total sugars. Sucrose levels are highly variable. Total anthocyanin pigment ranged from 4-1102 mg/L with a mean of 399. RED RASPBERRY ANTHOCYANIN PROFILES: Cultivars exhibit characteristic patterns distinguished by quantitative rather than qualitative differences. The profile can be altered by processing (enzymic changes) or storage conditions (partial hydrolysis). The HPLC analytical method was effective in detecting adulteration in 2 of 9 commercial concentrates. RED RASPBERRY PHENOLIC PROFILES: Phenolics were characterized and measured by HPLC/diode array techniques. While all samples were similar
qualitatively, they were quantitatively highly variable with respect to cultivar, maturity, mold and processing methodology. Mean total concentrations of flavonols and ellagic acid were 122 ppm and 28 ppm, respectively. Levels of quercetin and ellagic acid are of particular interest because of their possible beneficial role as anti-carcinogens.

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Progress 01/01/90 to 12/30/90

OutputsRED RASPBERRY JUICE COMPOSITION: Red raspberries were processed into juice for purposes of generating a compositional data base (n = 36) which incorporates the influences of variety, maturity, processing, mold contamination and geographical origin. Analysis of sugars, nonvolatile acids, anthocyanin pigments and phenolics are in progress. Methodology has been developed for analysis of the flavonoids. Ellagic acid which has been reported to have anticarcinogenic activity has been identified in red raspberry juice. Presence of substantial quantities of delphinidin glycosides in one commercial sample was evidence for adulteration. ROYAL OKANOGAN HUCKLEBERRY (ROH) JUICE: A number of acylated anthocyanin pigments have been identified in ROH juice, petunidin-3, 5-diglucoside acylated with p-coumaric acid being the major pigment. Acylation accounts for the intense purple color at pH 4.8. Pigment composition indicates that the fruit is the Garden Huckleberry (Solanum guineese)
rather than a Vaccinium species. CANTALOUPE JUICE CONCENTRATE: Clarified juice concentrate was produced from fresh and frozen fruits with and without rind. High yield and low acidity suggest its potential use as an alternative sweetener. Considerable ascorbic acid degradation occurred during processing. Juice concentrates showed very high nonenzymic browning rates during storage at 25(degree)C.

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Progress 01/01/89 to 12/30/89

OutputsCANTALOUPE JUICE: Juice concentrate was processed from fresh and frozen cantaloupe fruits, with and without rind. High juice yield (90%) and low acidity suggest its potential use as an alternate sweetener source. Compositional measurements in progress include sugar and nonvolatile acid profiles, total carotenoids, ascorbic acid and browning indices. Considerable ascorbic acid degradation occurs during processing. AMINO ACID DEGRADATION IN APPLE JUICE CONCENTRATES DURING STORAGE. HPLC analysis of apple juice concentrate stored for 9 months at 25C revealed significant losses of aspartic acid, asparagine, glutamine, arginine, threonine, methionine and isoleucine. Nonenzymic browning reactions are believed to account for much of these losses. RED RASPBERRY JUICE COMPOSITION: HPLC separation of red raspberry juice flavonoids has been achieved and peak identification is in progress. Compositional analysis of 25 authentic juice samples will include flavonoids, anthocyanin
pigments, sugars and nonvolatile acids. Characterization of red raspberry anthocyanin pigments in commercial concentrates is in progress. ROYAL OKANOGAN HUCKLEBERRY JUICE: Juice processed from Royal Okanogan Huckleberries had an intense purple color, high titratable acidity (0.79%) and high pH (4.8). Characterization of the anthocyanin pigments is in progress.

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Progress 01/01/88 to 12/30/88

OutputsAPPLE, PEAR AND WHITE GRAPE PHENOLICS: The flavonoids and cinnamates of apple, pear and white grape juice were characterized using HPLC-diode array detection and their changes during processing and storage measured. SO(subscript 2) addition protected cinnamates and procyanidins from oxidation, and blanching also had a protective effect on phenolics. Juice concentrate storage resulted in total loss of procyanidins and significant losses of cinnamates (c.a. 36%) and flavonoids (c.a. 50%). BROWNING OF JUICE CONCENTRATES DURING STORAGE: Apple, pear and grape juice concentrate exhibited zero order browning rate constants during storage at 25(degree)C. Initial browning values were not predictive of the final values or the browning rates. The formol values of juices proved to be a useful predictor of browning rates. CRANBERRY SUGAR, NONVOLATILE ACID AND ANTHOCYANIN PIGMENT PROFILES: The composition of Vaccinium macrocarpon, V. oxycoccus and V. vitis-idae were compared. While
these fruits showed different patterns, the differences in the sugars, acid and pigment profiles were quantitative rather than qualitative.

OutputsCRANBERRY JUICE COMPOSITION: Juice was processed from 20 lots of authentic cranberries and the following analyses performed: pH, titratable acidity, Brix, sugar profile, nonvolatile acid profile, anthocyanidin-anthocyanin pigment profile, anthocyanin pigment content, and color parameters. An HPLC system for analysis of anthocyanin pigments was developed which permitted quantitation of 6 anthocyanins and the tentative identification of 5 minor pigments. The results extend the base-line data for authentic cranberry juice and provide improved analytical methodology for detecting adulteration. APPLE JUICE COMPOSITION: Pattern recognition methods were able to classify authentic apple juice by variety and geographical origin from their glucose: fructose ratios and sucrose and sorbitol content. Pattern recognition based on sugar profiles was able to discriminate between potential adulterants and able to detect adulteration of apple juice at the 40% level. CHARACTERIZATION
OF COLORANTS: Analysis of grapeskin extract, elderberry extract, black currant, red cabbage, blackberry, and black raspberry juices are underway to provide indices for their detection in juices and beverages. Methods include HPLC determination of anthocyanidin and anthocyanin pigments in combination withspectral analysis.

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Progress 01/01/85 to 12/30/85

OutputsRED RASPBERRY JUICE COMPOSITION: Juice and concentrate were processed from 8 authentic berry samples. Sugar, nonvolatile acid, and pigment profiles were determined. HPLC methodology allowed the separation, identification, and quantitation of the major anthocyanin pigments. The distinctive pigment profile facilitates detection of added colorants. APPLE COMPOSITION: Commercial apple juice concentrate samples shipped from Argentina to the USA showed an increase in browning index and hydroxymethylfurfural content; there was no evidence for sucrose inversion or fumaric acid formation during shipping. Sugar, acid, and spectral profiles of the following potential apple juice adulterants were determined: invert beet sugar, corn syrup, malic acid, pear, pineapple, grape, raisin, fig, and plus juice concentrates. Pattern recognition multi-variate analysis is being applied to apple juice spiked with the adulterants to levels of 10, 20, and 40%. STRAWBERRY CELL-WALL
POLYSACCHARIDES: The compositional, sensory, and physical properties of fresh and frozen strawberries (Benton, Totem, and selva varieties) were compared. Selva had the highest percentage of acetone-insoluble solids, total pectin, and high molecular-weight polymers and its firmness was rated 2X and 1.3X that of Benton and Totem, respectively.

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Progress 01/01/84 to 12/30/84

OutputsCRANBERRY JUICE COMPOSITION & DETECTION OF ADULTERATION IN COMMERCIAL CRANBERRY DRINK & CONCENTRATE--Cranberries were processed into juice and concentrate and the non-volatile acid, sugar, and anthocyanidin profiles determined by HPLC. Anthocyanin pigement concentration and Hunter color parameters were also determined. Analysis of 22 commercial samples showed 13 to be adulterated with added citric and/or malic acid and a natural colorant believed to be grape-skin extract. APPLE JUICE COMPOSITION--Sugar, non-volatile acid, phenolic and u.v.-visible spectrophotometric profiles were determined for apple juice prepared from 8 authentic samples of USA and international origin. The sugar and acid composition agreed with values previously reported but there was evidence for reduced sucrose and sorbitol content as result of postharvest storage. Substantial quantities of quinic acid was found in most samples. Processing (particularly gelatin fining) had a major influence
on the phenolic profile. RED RASPBERRY JUICE COMPOSITION--Juice and concentrate was produced from the major varieties grown in the Pacific Northwest. HPLC is being used for determining the sugar, non-volatile acid and anthocyanidin profiles. VARIETY & MATURITY INFLUENCE ON STRAWBERRY PECTIC SUBSTANCES-Extraction and fractionation procedures have been developed for characterizing the water soluble, chelator soluble, and insoluble fractions.

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Publications

Progress 01/01/83 to 12/30/83

OutputsAPPLE JUICE COMPOSITION--Juice was processed from authentic apple samples. Compositional analyses included PGDTC/PGyC ratios, u.v.-visible spectra and non-volatile acids. The DEL PGDTC was considerably lower for the acid fraction (-27.6) than for the sugars (-23.2); there were no distinct differences for DEL PGDTC values that could be attributed to variety or geographic origin. There was considerable variation in quinic acid content; shikimic acid was identified in minor amounts. CRANBERRY JUICE COMPOSITION.--The following analyses were carried out on commercial cranberry concentrates and drinks: u.s.-visible spectra, anthocyanin content, nonvolatile acids by HPLC, and anthocyanidins by HPLC. The latter method shows promise for detection of grape-skin extract as an adulterant. SAPONINS IN FRUITS--Blackberry, strawberry and plum fruit extracts were tested for their ability to form a stable foam, induce hemolysis of erythrocytes, and produce colors characteristic of
triterpenes or sterols. There is strong evidence for presence of saponins in blackberries. Strawberries contain compounds which will produce a stable foam, but the presence of saponins is not substantiated. There is no evidence for saponins in plum fruit. INFLUENCE OF MOLD CONTAMINATION AND ULTRAFILTRATION ON COLOR STABILITY OF STRAWBERRY JUICE 7 CONCENTRATE--Juices made from moldy strawberries were clarified by conventional filtration and ultrafiltration and processed into concentrates.

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Publications

Progress 01/01/82 to 12/30/82

OutputsSTABLE CARBON ISOTOPE ANALYSIS OF APPLE COMPONENTS--Apple juice and concentrate was processed in our pilot plant and 1 3/C 1 2/C analyses were performed on the following fractions: whole apple pulp, apple juice, apple seeds, apple sugars, and apple non-volatile acids. The del 1 3/C of the acid fraction is considerably higher than the sugars; thus variation in total acidity may account for del 1 3/C variation among different apple samples. STRAWBERRY WINE COLOR--Color and composition of strawberry wines made from ripe, overripe, and mold-contaminated fruit (Benton and Totem varieties) were monitored during storage. Overripe fruit with its higher anthocyanin and total phenolics gave wines with better color. Totem fruit had higher anthocyanin, lower polyphenoloxidase activity and lower total phenolics and produced wines with better color. Mold contamination increased juice viscosity, reduced fermentation rate, and accelerated color degradation. Anthocyanin content
and Hunter L values of wines were highly correlated with panel color evaluations. SAPONINS IN FRUITS--Blackberries, strawberries and plums were analyzed for saponins. Use of PVPP was effective in removing phenolics which interfere with detection and chromatography of saponins. Both blackberries and strawberries contain saponins; evidence for their presence is based on reactivity with diagnostic reagents and foaming and hemolytic activity.

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Publications

Progress 01/01/81 to 12/30/81

OutputsThe sugar and non-volatile acid composition of strawberries as related to maturity have been compared utilizing High Performance Liquid Chromatography (HPLC), Gas Liquid Chromatography (GLC), and enzymic procedures. Sucrose and malic acid decreased greatly in overripe fruit. Good agreement was obtained for the different methods, HPLC offering the advantages of speed and versatility. Percent sorbitol content was found to be a useful index for detecting sucrose addition in frozen cherries and juice concentrates derived from the sugar-packed fruit. Native invertase caused sucrose inversion in all samples. Work was initiated on the influence of maturity and mold contamination on the color quality of strawberry wine. Compositional analyses of ripe and overripe fruit (Benton and Totem varieties) included: anthocyanin pigment, leucoanthocyanins, flavanols, total phenolics, ascorbic acid, polyphenol oxidase, and Beta-glucosidase. Wines will be made from the fruit and the
color quality evaluated. Evaluations will also be made on wines derived from mold-contaminated fruit.

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Publications

Progress 01/01/80 to 12/30/80

OutputsThe glucose, fructose, sucrose, and sorbitol contents of fruits of economic importance in the Northwest were compiled from the literature and analyzed statistically. The different fruits show characteristics sugar patterns with only moderate variation considering the diversity of geographic origin, variety, maturity, and method of analysis. This data provides a useful baseline for comparing the sugar composition of analyzed samples in screening for authenticity. The sugar and non-volatile acid composition of strawberries as related to maturity have been investigated. The sugar and sorbitol contents of cherries have been analyzed to determine whether addition of sucrose to commercial samples could be determined. Sucrose inversion occurred in all samples but percent sorbitol content could be used to detect sucrose addition. Sorbitol content is a useful index for detecting adulteration of non-sorbitol containing fruits such as blackberry, strawberry, and raspberry
with cheaper sorbitol-containing fruits such as pear, apple, and plum. Analysis of wines made from sorbitol-containing blackberry concentrates showed that sorbitol was not fermented and suggested that its presence in blackberry wines could be used as an index of adulteration.

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Publications

Progress 07/01/79 to 12/30/79

OutputsThe sugar and organic acid composition of 15 blackberry samples has been reported. Glucose and fructose are the only sugars present, the glucose: fructose ratio for all samples being 0.710. The acid composition shows more complexity and variation, lacto-isocitric being the major acid in some samples and absent in others. Sugar composition can be a useful screening criteria for determining the authenticity of fruit products. Current research methods (HPLC) and to the sugar and acid composition of other fruits -- cherry, pear, strawberry. Browning of pear juice concentrate during storage can be effectively inhibited by treatment of juice with cation exchange resin to remove amino acids. It was concluded that Maillard browning reactions predominated during storage.